| ObjectPhotoaging is a premature skin aging process, due to continuously long time exposure to ultraviolet radiation of sun and characterized by clinical features, such as Sunburn, coarse wrinkles, and sex keratosis to lose elasticity and light. About more than 80% of the exposed parts cosmetic problems is caused by skin photoaging, leading to serious consequences. Photoaging was found closely related to the occurrence of a variety of skin diseases, such as actinic elastosis, solar sex keratosis, chronic actinic cheilitis, sun freckles blue ubber-bleb nevus, basal cell carcinoma (BCC), squamous cell carcinoma, melanin cell carcinoma, etc, and even skin cancer. The study of photoaging has been performed extensively both here and abroad in the past decades. A large number of literature were consulted and the molecular mechanism way was map, so we can found the occurrence of skin photoaging mechanism is related to oxidative stress and inflammation factors.Patchouli as one of Aromatic herbs eliminating dampness, with the effect of eliminating dampness with aromatics, alleviating vomiting by regulating stomach, and clearing summerheat by relieving superficies syndrome, is commonly used to therapy syndromes of dampness accumulate in stomach and to therapy syndromes of vomiting caused by gastric cavity-ruffian flatulence in China. It was commonly used in facial beauty fomula in Chinese ancient prescriptions, can also be used to treat facial diseases.Patchouli oil extracted by steam distillation from Patchouli is the main medicinal parts of the patchouli. It has become one of the most important ingredients in cosmetics because of its medica characteristics and stability properties. It contains a variety of biological active ingredient, has the activity of anti-allergic, antimicrobial, antioxidant and anti-inflammatory, et al. The path of Patchouli oil developed antioxidant and anti-inflammatory activity are associated with its anti-photoaging activity. Is there any relationship between the facial beauty using of patchouli oil and anti-phontoaging activity? Does Patchouli oil have the activity of anti-phontoaging? What is the machanism?Therefore, patchouli oil was chosen to observed the protection effect against photoaging caused by ultraviolet in mice. The possible mechanism was explored by histopathological method and through oxidative stress path.Methods and results1. Chemical composition analysis of Patchouli oilGas chromatograph was used to determine the content of Patchouli alcohol, the main chemical indicators in patchouli oil. GC-MS instrument was used to determine the chemical composition of patchouli oil. The measured content of Pachouli alcohol is 28.2% (g/g), accorded with the requirement of Chinese pharmacopoeia of no less than 26%. 18 kinds of chemical compositions were identified, accounted for 88.3% of total volatile oil. Terpenoids is the main composition. Monoterpene and Sesquiterpene together accounts for above 50% and Patchouli alcohol accounts for 29.3% (peak area percentage). The quality of samples used for pharmacodynamics experiment could be ensured by qualitative identification and quantitative analysis.2. The effect of Patchouli oil to photoaging in mice63 KM mice, used to be experimental animals, were randomly divided into seven groups, such as NC, SC, MC, VC, PO-H, PO-M, and PO-L group. Each group has 9 mice. UVA with UVB were used to induce skin photoaging Model. Patchouli oil was applied on the bare skin before UV radiation. In the experiment, dorsal skins were shaved after the mice were anesthetized by ether inhalation. In two hours before UV irradiation, the shaved dorsal skins of mice were applied topically with Patchouli oil. Macroscopic score evaluation and skin elasticity test with the index of macroscopic score and the recovery time of skin were used to estimate the protect effect of patchouli oil on photoaging model in mice. According to the result of macroscopic score, application with Patchouli oil at high dose (PO-H group) had a tendency to suppress the UV-induced extensive erythemas and deep wrinkles after six weeks treatment; In addition, the PO-M group did not exhibit its inhibited effect on skin wrinkle formation until the eighth week. After 10 weeks, the scores of the PO-H group and PO-M group were significantly lower than that of the VC group, indicating that topic application of PO prevented the UV-induced macroscopic skin damages. According to the result of skin elasticity test, although the PO-L group did not reduce the recovery time significantly as compared to the VC group, the recovery times of the mice in the PO-M and PO-H groups were significantly shorter than that in the VC group. These results demonstrated that treatment with Patchouli oil (especially 6-9 mg/mouse) could promote the skin elasticity. The result of macroscopic score evaluation and skin elasticity test jointly indicated PO applied topically could protect against photoaging model induced by UV radiation in mice.3. The mechanism of patchouli oil protect against skin photoagingHE staining and Gomori’s aldehyde fuchsin staining were used to observe the effects of patchouli oil on organization structure in aging skin, by obseving the change of thickness of Epidermis and structure of cuticle, the form, quantity and distribution of Collagen fiber and Elastic fibers. According to the results of Epidermis thickness determination, PO-M group and PO-H group show thinner Epidermis obviously, decrease 19.8μm and 23.7μm respectively compared with the VC group. Results suggest the PO-M, the PO-H group can significantly reduce the epidermal hyperplasia induced by UV. The results of observation of the organizational characteristics indicated patchouli oil applied topically could improve the abnormally changes induced by UV in skin structure, such as abnormal thickening of the epidermis, flattening of DEJ, disappearing of cuticular process and corpora papillare, crack, decrese and arranged in disorder of Collagen fiber and denaturation and crack of elastic fibers.The cotent of collagen and malondialdehyde (MDA), and activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and catalase (CAT) were determined by corresponding Biological reagent kit to investigate the mechanism. The results shows, compared with VC group, PO-M and PO-H group can significantly improve the content of collagen by 21.9% and 26.3% respectively, significantly reduce the amount of MDA, also increase the activity of SOD and GSH-Px significantly. PO-L and PO-H group significantly increased CAT activity by 9.8% and 90.7% respectively. PO-M group did not have significant effects on the CAT activity. Results suggest that patchouli oil can inhibit the damage of collagen, the reaction of lipid peroxidation, and the activity decreasing of SOD, GSH-Px and CAT,both induced by UV in mice.4. PO playing the efficacy of dampness elimination and heat celaring to protect against photoaging was explained with modern science.Combined with the molecular mechanism and the theory of traditional Chinese medicine, the modern scientific connotation was discussed. UV irradiation is "heat" and ROS belong to the "phlegm". Photoaging induced by Ultraviolet can be explained with photoaing caused by external heat and skin cell dysfunction caused by ROS damage can be explained with kin blood circulation disorders caused by phlegm stasis s in the theory of Chinese medicine. PO inhibiting skin abnormal structure change induced by Ultraviolet can be explained with clearing summerheat by relieving superficies syndrome. PO inhibiting ROS damage with aromatic properties can be explained with eliminating dampness with aromatics;Therefore, PO plays the efficacy of dampness elimination and heat celaring to protect against photoaging.ConelusionAccording to the understanding of enzymatic antioxidant defense system can regulate the ROS scavenging free radicals, antioxidant therapy is considered to be a effective way to resist skin photoaging. Patchouli oil as an antioxidant, applied topically can protect against photoaging caused by UV radiation in mice. This effect may be developed by reducing hyperplasia of epidermis, restraining the change of the skin organization structure and inhibiting collagen damage. Inhibiting lipid peroxidation and increasing the activity of antioxidant enzymes, related to its inhibition mechanism of ROS damage induced by UV, may be the mechanism. This is the first study comprehensively proof the protect effect of patchouli oil on photoaging in mice. It can provide patchouli oil an evidence to be used as a potentially cosmetic product for skin care. The modern scientific connotation of PO playing efficacy of dampness elimination and heat celaring to protect against photoaging were explained.
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